Timely and accurate estimation of Above-Ground-Biomass (AGB) in cotton is essential for precise production monitoring. The study was conducted in Shaya County, Aksu Region, Xinjiang, China. It employed an unmanned aerial vehicle (UAV) as a low-altitude monitoring platform to capture multispectral images of the cotton canopy. Subsequently, spectral features and textural features were extracted, and feature selection was conducted using Pearson’s correlation (P), Principal Component Analysis (PCA), Multivariate Stepwise Regression (MSR), and the ReliefF algorithm (RfF), combined with the machine learning algorithm to construct an estimation model of cotton AGB. The results indicate a high consistency between the mean (MEA) and the corresponding spectral bands in textural features with the AGB correlation. Moreover, spectral and textural feature fusion proved to be more stable than models utilizing single spectral features or textural features alone. Both the RfF algorithm and ANN model demonstrated optimization effects on features, and their combination effectively reduced the data redundancy while improving the model performance. The RfF-ANN-AGB model constructed based on the spectral and textural features fusion worked better, and using the features SIPI2, RESR, G_COR, and RE_DIS, exhibited the best performance, achieving a test sets R2 of 0.86, RMSE of 0.23 kg·m−2, MAE of 0.16 kg·m−2, and nRMSE of 0.39. The findings offer a comprehensive modeling strategy for the precise and rapid estimation of cotton AGB.